大鼠骨髓间充质干细胞对减体积肝移植大鼠的作用及机制
The role and mechanism of BMMSCs in reduced-size liver transplantation of rats
摘要目的:研究骨髓间充质干细胞(BMMSCs)对大鼠减体积移植肝作用及机制。方法:选取体重210~250 g的健康雄性清洁级Lewis大鼠和Brown Norway (BN)大鼠各40只,分别作为供体和受体建立大鼠50%减体积肝移植免疫排斥模型。模型大鼠分为干细胞组( n=20)和生理盐水组( n=20)。以体重80~100 g的健康雄性BN大鼠制备BMMSCs,经阴茎背静脉注射到肝移植受体。于术后即刻、第1天、第3天、第7天分别取5只大鼠肝组织,进行组织病理学观察和移植排斥反应评分,并利用免疫组织化学染色和蛋白质电泳检测移植肝微管相关蛋白1轻链3(LC3)和自噬调控蛋白Beclin-1的表达。 结果:生理盐水组肝移植受体大鼠术后第1、3、7天的急性排斥反应评分分别为(2.33±0.58)分、(4.00±0.00)分、(6.33±0.58)分,干细胞组分别为(2.10±0.58)分、(3.73±0.58)分、(5.67±1.15)分,两组比较呈降低趋势,差异有统计学意义( P<0.05)。与生理盐水组比较,在术后第1、3、7天干细胞组肝组织自噬相关蛋白LC3和Beclin-1表达量增多,差异有统计学意义( P<0.05)。 结论:BMMSCs移植对大鼠减体积肝移植术后的移植肝具有保护作用,自噬在其中发挥一定作用。
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abstractsObjective:To investigate the effect and mechanism of bone marrow mesenchymal stem cells (BMMSCs) on liver transplantation with 50% reduced-size in rat models.Methods:For 40 normal male Brown Norway(BN) and Lewis rats weighing 210-250 g were included respectively to generate the acute rejection models following 50% reduced-size liver transplantation in rats. The recipients were divided into BMMSCs group ( n=20) and normal saline group ( n=20). Healthy male BN rats were used to prepare BMMSCs. Transplanted liver tissues were collected at 0h, 1d, 3d, 7d post the transplantation for further analysis. Pathological changes and the extent of rejection were evaluated under the light microscope. The levels of microtubule-associated protein 1 light 3 (LC3) and autophagy regulator Beclin-1 proteins were detected by immunohistochemical analysis and Western blotting. Results:Rejection activity indices of the normal saline group at 0d, 3d, 7d after the surgery were (2.33±0.58), (4.00±0.00), (6.33±0.58). The BMMSCs group were (2.10±0.58), (3.73±0.58), (5.67±1.15), which was decreased comparing with normal saline group, difference was statistically significant ( P<0.05). On the 1d, 3d, 7d after the transplantation, compared with normal saline group, expression of autophagy-related proteins LC3 and Beclin-1 in BMMSCs group was increased ( P<0.05). Conclusions:It showed that autophagy has an effect on the protection of BMMSCs liver graft of rats.
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